甜高粱蔗糖积累关键时期叶片转录组测序分析  

Transcriptome Sequencing Analysis of Sweet Sorghum Leaves in the Critical Period of Sucrose Accumulation

作　　者：陈思孛 赵质涵 马爱军 张敖[1] 王志和 阮燕晔[1] CHEN Si-bo;ZHAO Zhi-han;MA Ai-jun;ZHANG Ao;WANG Zhi-he;RUAN Yan-ye(Biological Science ＆ Technology College, Shenyang Agricultural University, Shenyang 110866;Liaoning Denghai Seed Co. Ltd., Shenyang 110000)

机构地区：[1]沈阳农业大学生物科学技术学院,沈阳110866 [2]辽宁登海种业有限公司,沈阳110000

出　　处：《生物技术通报》2018年第3期98-104,共7页Biotechnology Bulletin

基　　金：辽宁省科学计划(2015103001)

摘　　要：通过转录组测序技术,在甜高粱糖分累积的关键时期寻找甜高粱和普通高粱组织间差异表达的相关基因,通过相关基因的功能注释及相关联的Pathway代谢通路分析,进一步筛选与糖分积累与代谢相关的基因,以利于为充分挖掘甜高粱的生物学潜能,改良高粱品种提高其茎杆的糖含量打下坚实基础。以甜高粱辽甜1号和粒用高粱新苏2号为材料,对甜高粱与粒用高粱在成熟期进行叶片转录组数据测序分析并比较,测序结果如下,共得到71.98 M条reads,总碱基数14.54 Gb。使用Cufflinks软件对reads进行组装,共发掘出1 562个新基因。通过差异表达筛选出3 115个差异基因,其中1 499个上调,1 616个下调。对差异基因进行GO、COG分类和KEGG代谢途径富集性分析,将这些差异表达基因归类于包括淀粉与蔗糖代谢途径在内的94个代谢途径,从淀粉与蔗糖代谢通路(Pathway ID:Ko00500)中取β-呋喃果糖苷酶、蔗糖合成酶、果糖激酶的5个差异基因分别为Sb06g031910、Sb06g023760、Sb01g035890、Sb01g033060和Sb10g0082805进行实时荧光定量分析,发现这些基因在不同生育时期的表达与其对应酶类的活性有紧密的联系,推测这些基因在甜高粱叶片的蔗糖合成及积累过程中发挥着至关重要的作用。The aim of this study is to find the genes that differentially express between sweet sorghum and common sorghum in the criticalperiod of sweet sorghum sugar accumulation through transcriptome sequencing. Functional analysis of related genes and associated Pathwaymetabolic pathways were used to further screen the genes related to sugar accumulation and metabolism,which is for laying a solid foundationin fully exploring the biological potential of sweet sorghum and improving sorghum varieties to increase the sugar contents of their stems. In thisstudy,sweet sorghum Liaotian 1 and grain sorghum Xinsu 2 were used as research materials,and their leaf transcriptome data were sequencedand compared at maturity. The 71.98 M reads and 14.54 G total bases were obtained by sequencing. The Cufflinks software was used to assemblethe reads,and 1562 new genes were discovered compared with known genetic model. Through differentiated expression,3 115 expresseddifferential genes were screened,of which 1499 were up-regulated and 1616 were down-regulated. Through GO,COG classification andKEGG pathway enrichment analysis,all of these differentially expressed genes were classified into 94 metabolism pathways covering starch andsucrose metabolism. The five differential genes of β-fructofuranosidase,sucrose synthase and fructokinase from the starch and sucrose metabolicpathway（Pathway ID ：Ko00500）were respectively Sb06g031910,Sb06g023760,Sb01g035890,Sb01g033060,and Sb10g0082805 forreal-time fluorescence quantitative analysis. The results showed that the expression of these genes at different growth stages was closely related tothe activity of their corresponding enzymes,inferring that these genes play a crucial role in the synthesis and accumulation of sucrose in sweetsorghum.

关 键 词：甜高粱 成熟期叶片 转录组 糖分积累与代谢 蔗糖 

分 类 号：S566.5[农业科学—作物学]